GloLIVE Human/Mouse SSEA-4 NL557 Live Cell Antibody

Catalog #: NLLC1435R Datasheet / COA / SDS
50X Concentration, Azide Free
Catalog # Availability Size / Price Qty
NLLC1435R
Detection of SSEA-4 in Live BG01V Human Embryonic Stem Cells.
1 Image
Product Details
Procedure
FAQs
Reviews

GloLIVE Human/Mouse SSEA-4 NL557 Live Cell Antibody Summary

Kit Summary

To detect SSEA-4 expression in live cells by immunocytochemical staining.

 

Why confirm marker expression in live cells before colony selection?

Maintaining pluripotent human embryonic stem cells and deriving induced pluripotent stem cells require colony selection and cell expansion. Picking colonies that contain pluripotent stem cells is typically achieved by manually analyzing colony morphology. Unfortunately, this critical process is time-consuming and labor-intensive.

In addition, morphology-based colony selection does not consistently guarantee pluripotent starting populations, since cells can fail to be fully reprogrammed or can naturally differentiate within a colony.

To address this need, R&D Systems offers GloLIVE™ antibodies that allow researchers to confirm stem cell marker expression in live cells before colony selection.

Live pluripotent stem cell imaging:

  • Promotes the selection of high quality, undifferentiated stem cell colonies to reduce experimental variation.
  • Verifies stem cell pluripotency in 30 minutes, ensuring efficient use of time and reagents.
  • Allows for the continued culture of pluripotent cells with no adverse effects on cell proliferation or stemness following staining.

 

Antibody Components
  • GloLIVE NL557-conjugated Mouse Anti-Human/Mouse SSEA-4 Monoclonal Antibody.
  • Supplied as 50X concentration of antibody in 0.5 mL PBS

Azide free and endotoxin tested (= 5 EU/mL), this antibody can be used for 12 tests, if a staining volume of 2 mL is used.

Specifications

Shipping Conditions
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8° C. Do not use past expiration date. Protect from light.
Applications
Immunocytochemistry
Species
Human Mouse

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Product Datasheets

You must select a language.

x

Scientific Data

Detection of SSEA-4 in Live BG01V Human Embryonic Stem Cells. The stem cell marker SSEA-4 was visualized in live BG01V human embryonic stem cells using a GloLIVE NL557-conjugated Mouse Anti-Human/Mouse SSEA-4 Monoclonal Antibody (Catalog # NLLC1435R; red). The cells were counterstained with Hoechst 33342 (blue). Positive staining for SSEA-4 in human stem cells is an indicator of pluripotency.

Background: SSEA-4

SSEA-4 is expressed on the surface of human embryonal carcinoma (EC) cells (the pluripotent stem cells of teratocarcinomas), human embryonic germ cells (EG), and human embryonic stem cells (ES). Expression of SSEA-4 is down-regulated following differentiation of human EC cells. In contrast, the differentiation of murine EC and ES cells may be accompanied by an increase in SSEA-4 expression (1-4).

References
  1. Shevinsky, L.H. et al. (1982) Cell 30:697.
  2. Kannagi, R. et al. (1983) EMBO J. 2:2355.
  3. Thomson, J.A. and J.S. Odorico (2000) Trends Biotechnol. 18:53.
  4. Draper, J.S. et al. (2002) J. Anat. 200:249.
Long Name
Stage-specific Embryonic Antigen-4
Alternate Names
SSEA4; SSEA-4; SSEA-4;stage-specific embryonic antigen 4

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, SSEA-4 expression can be verified in live cells prior to colony selection or experimentation using this simple procedure:

  • Add the SSEA-4 antibody directly to the media of cultured cells
  • Wash the cells after a 30 minute incubation
  • Select for and expand SSEA-4-positive colonies
 
 

Precautions

Use aseptic technique and sterile culturing conditions to prevent contamination.

Reagents Provided

GloLIVE NL557-conjugated Mouse Anti-Human/Mouse SSEA-4 Monoclonal Antibody. (Catalog # NLLC1435R):

Other Supplies Required

Reagents

  • Stem cell culture media

Materials

  • Human or mouse pluripotent stem cells
  • Pipettes and pipette tips
  • Serological pipette

Equipment

  • 37 °C and 5% CO2 incubator
  • Fluorescence microscope
 

Live Cell Immunocytochemistry Procedure Overview

To ensure sterility of cultures, all steps should be performed under sterile conditions.


Add primary antibody in appropriate culture media to cells.
 
  1. Add fluorochrome-conjugated primary antibody in appropriate culture media to cells.
  2. Incubate for 30 minutes.

Replace with fresh culture media
 
  1. Replace with fresh culture media.

Visualize using a fluorescence microscope
 
  1. Visualize using a fluorescence microscope.

Note: Culture with GloLIVE antibodies does not appear to affect cell proliferation or stemness as assayed by proliferation curves for 3 days post-antibody incubation and expression levels of SSEA-4 and Oct-3/4.

FAQs

  1. What is the recommended starting confluence to observe staining?

    • The optimal confluence would depend on the purpose of the staining. If the plan is to use all the cells in the flask after they have proliferated, then one can wait for the cells to become confluent or almost confluent (80-90% confluent) and then check the staining. If the goal is to to check on the cells mid-way through their growth and there are sufficient cells in the flask, one could check the staining then as well. The staining is not dependent on how many cells are in the flask, as long as there are sufficient cells.

View all Antibody FAQs

Reviews for GloLIVE Human/Mouse SSEA-4 NL557 Live Cell Antibody

There are currently no reviews for this product. Be the first to review GloLIVE Human/Mouse SSEA-4 NL557 Live Cell Antibody and earn rewards!

Have you used GloLIVE Human/Mouse SSEA-4 NL557 Live Cell Antibody?

Submit a review and receive an Amazon gift card.

$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image

$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image

Submit a Review